Method for extracting superoxide dismutase from cattle and sheep blood largly industrially and blade type agitator applied to same

A superoxide, large-scale technology, applied in mixer accessories, chemical instruments and methods, biochemical equipment and methods, etc., can solve the problems of long temperature gradient processing time, large equipment investment, long heating time, etc., and achieve uniform temperature. And fast, reduce process cost, and is conducive to the effect of removal

Inactive Publication Date: 2011-07-06
云南光华亚南生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] A. Cu-Zn-SOD is easily denatured in the presence of organic solvents such as ethanol and chloroform, which will promote the conformational change of the egg self-quality
[0009] B. Organic solvents will cause about 8% loss of copper ions, and if the metal prosthetic group is removed, its stability will be greatly reduced, affecting the enzyme activity of SOD
[0010] C. This method consumes a large amount of organic reagents, increases production costs, and pollutes the environment
[0011] D. Expensive column chromatography system, column packing and high-speed centrifuge. The equipment investment is large, and after column chromatography, the yield of the product is greatly reduced, the ratio of input to output is extremely uneconomical, and large-scale chemical production cannot be realized
[0012] Even in the large-scale industrial production that does not use organic solvents to extract SOD, due to the heating in a large container, the difficulty of the direct thermal denaturation method is "uneven heating", and the temperature gradient treatment time for the supernatant is too long For example, there are some 6-level temperature gradient treatments with a total time of nearly 200 hours. Due to the long heating time, this kind of treatment not only easily leads to the denaturation of the target protein to be extracted, but also cannot "rapidly" heat up and down. Effective removal of foreign proteins, resulting in a decrease in the yield and specific activity of the target protein

Method used

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  • Method for extracting superoxide dismutase from cattle and sheep blood largly industrially and blade type agitator applied to same
  • Method for extracting superoxide dismutase from cattle and sheep blood largly industrially and blade type agitator applied to same
  • Method for extracting superoxide dismutase from cattle and sheep blood largly industrially and blade type agitator applied to same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Example 1: Large-scale industrialized method for extracting superoxide dismutase from cattle blood

[0092] 1. Add 30g of sodium citrate anticoagulant to 10Kg of fresh bovine blood. Let it stand at a low temperature of 4°C for 30 minutes. After the solution is separated, remove most of the supernatant.

[0093] 2. Add the same volume of distilled water, stir, hypotonic hemolysis for 30 minutes, and then stand for 40 minutes. After the solution is separated, the supernatant is ultrafiltered (using a polyacrylonitrile membrane with a pore size of 1μm) to obtain 2000ml of supernatant Liquid and 6500ml precipitation;

[0094] While heating the precipitate, stir it with a mixer with a heat source to quickly heat it to 40°C within 8 minutes. After keeping it for 20 minutes, stir with a mixer with a cold source while reducing the temperature to quickly reduce the temperature to within 6 minutes. After centrifugation at 4000 rpm for 20 minutes at room temperature, let it stand for 2...

Embodiment 2

[0099] Example 2: Large-scale industrialized method for extracting superoxide dismutase from cattle blood

[0100] 1. Add 50g of sodium citrate anticoagulant to 10Kg of fresh bovine blood. Let it stand for 50 minutes at a low temperature of 4°C. After the solution is separated, remove most of the supernatant.

[0101] 2. Add the same volume of distilled water, stir, hypotonic hemolysis for 30 minutes, and then leave for 50 minutes. After the solution is separated, the supernatant is ultrafiltered (using a polyacrylonitrile membrane with a pore size of 5μm) to obtain 2500ml of supernatant Solution and 6000ml precipitation;

[0102] While heating the precipitate, stir it with a mixer with a heat source to quickly heat it to 45°C within 8 minutes. After holding for 10 minutes, stir with a mixer with a cold source while reducing the temperature to quickly reduce the temperature in 8 minutes. After centrifugation at 5,000 rpm for 15 minutes at room temperature, let it stand for 10 minut...

Embodiment 3

[0107] Example 3: Large-scale industrialized method for extracting superoxide dismutase from cattle blood

[0108] 1. Add 60g of sodium citrate anticoagulant to 10Kg of fresh bovine blood. Let it stand for 60 minutes at a low temperature of 4°C. After the solution is separated, remove most of the supernatant.

[0109] 2. Add the same volume of distilled water, stir, hypotonic hemolysis for 30 minutes, and then stand for 60 minutes. After the solution is separated, the supernatant is ultrafiltered (using a polyacrylonitrile membrane with a pore size of 5μm) to obtain 3500ml of supernatant Liquid and 5800ml precipitation;

[0110] While heating the precipitate, stir it with a mixer with a heat source to quickly heat it to 55°C within 10 minutes. After keeping it for 30 minutes, stir it with a mixer with a cold source while cooling down to quickly reduce the temperature within 10 minutes. After centrifugation at 6000 rpm for 20 minutes at room temperature, the precipitate was removed ...

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Abstract

The invention discloses a method for extracting superoxide dismutase from cattle and sheep blood largly industrially, comprising the following steps of: carrying out the production steps of anticoagulation, hypotonic hemolysis, ultrafiltration, fast temperature rise and reduction, sedimentation, three-stage temperature treatment, salting-out, ultrafiltration, concentration, freeze drying, and thelike on fresh cattle or sheep blood so as to simultaneously obtain two groups of superoxide dismutase finished products which are slightly different in specific activity and purity, wherein the fast temperature rise and reduction is realized through an agitator with a heat / cold source; the first-stage temperature treatment is carried out at 45-55 DEG C, the second-stage temperature treatment is carried out at 55-60 DEG C, and the third-stage temperature treatment is carried out at 60-65 DEG C. Compared with the prior art, the invention has the advantages of simple process, short production period, high yield coefficient and specific activity of the products, stable quality, low cost, excellent economic benefit, and the like.

Description

Technical field [0001] The invention relates to a method for extracting superoxide dismutase, in particular to a large-scale industrialized method for extracting superoxide dismutase from the blood of cattle and sheep. Background technique [0002] Superoxide dismutase (SOD) is a metal enzyme that can catalyze the disproportionation of superoxide anion radicals. It is an extremely effective antioxidant that can resist the damage of superoxide anion radicals to cells. Studies have shown that SOD may be one of the enzymes that collapse the leek in the human body. SOD can be divided into two groups structurally, Cu-Zn-SOD is the first group, Mn-SOD and Fe-SOD are the second group, and Cu- Zn-SOD is a kind of SOD that people know the most, and it is a breakthrough in the research of SOD structure. With the in-depth expansion of the application of SOD in clinical medicine, food and cosmetics, the demand for SOD is increasing. The preparation of SOD from cattle blood will move from la...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/08B01F7/00B01F15/06
Inventor 李蒙
Owner 云南光华亚南生物科技有限公司
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